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Duplex real-time PCR assay for the simultaneous detection of Ophiostoma novo-ulmi and Geosmithia spp. in elm wood and insect vectors

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Duplex real-time PCR assay for the simultaneous detection of Ophiostoma novo-ulmi and Geosmithia spp. in elm wood and insect vectors

by
Pepori, Alessia L.; Luchi, Nicola; Pecori, Francesco; Santini, Alberto

Publication date
Usage
Attribution 4.0 InternationalCreative Commons Licenseby
Topics
DNA quantification, duplex qPCR, Dutch Elm Disease, Geosmithia spp. life cycle, Ophiostoma novo-ulmi, Scolytus multistriatus
Publisher
Pensoft Publishers
Collection
biodiversity
Contributor
Pensoft Publishers
Language
English
Rights
https://biodiversitylibrary.org/permissions
Rights-holder
Copyright held by individual article author(s).
Volume
84
Item Size
21.8M
Abstract
Dutch elm disease (DED) is a destructive tracheomycosis caused byOphiostoma novo-ulmi, an ascomycete probably originating in East-Asia that is devastating natural elm populations throughout Europe, North America and Asia. The fungus is mainly spread by elm bark beetles that complete their life cycle between healthy and diseased elms. Recently, it has been highlighted that some fungi of the genusGeosmithia, which are similarly well associated with bark beetles, seem to also play a role in the DED pathosystem acting as mycoparasites ofO. novo-ulmi. Although some relationship between the fungi is clear, the biological cycle ofGeosmithiaspp. within the DED cycle is still partly unclear, as is the role ofGeosmithiaspp. in association with the bark beetles. In this work, we tried to clarify these aspects by developing a qPCR duplex TaqMan assay to detect and quantify DNA of both fungi. The assay is extremely sensitive showing a limit of detection as low as 2 fg μl–1for both fungi. We collected woody samples from healthy and infected elm trees throughout the beetle life cycle. All healthy elm samples were negative for bothGeosmithiaspp. andO. novo-ulmiDNA.Geosmithiaspp. are never present in infected, but living trees, while they are present in frass of elm bark beetles (EBB –Scolytusspp.) and at each stage of the EBB life cycle in much higher quantities thanO. novo-ulmi. This work provides a better understanding of the role and interactions occurring amongst the main players of the DED pathosystem.
Addeddate
2025-04-01 17:15:58
Bhl_virtual_titleid
210923
Bhl_virtual_volume
v.84 (2023)
Call number
10_3897_neobiota_84_90843
Call-number
10_3897_neobiota_84_90843
Foldoutcount
0
Genre
article
Identifier
duplexrealtimep84pepo
Identifier-ark
ark:/13960/s299c1rvtk9
Identifier-bib
10_3897_neobiota_84_90843
Identifier-doi
10.3897/neobiota.84.90843


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